7 results about "Cell dissociation" patented technology

This invention provides a method for propagating pluripotent stem cells (PSCs) by suspension culture in a bioreactor. [Solution] The method comprises the following steps: (i) adding a ROCK inhibitor (ROCKi) to pluripotent stem cells cultured in suspension in a bioreactor; (ii) adding a cell dissociation agent, thereby dissociating aggregates of pluripotent stem cells; (iii) diluting the cell dissociation agent added in step (ii) by adding a sufficient excess volume of culture medium to reduce the concentration of the cell dissociation agent to a concentration at which cell aggregates can be re-formed; and (iv) culturing the mixture obtained in step (iii) under appropriate conditions that allow for the proliferation of PSCs.

The invention discloses an efficient universal mouse pancreatic tissue single-cell suspension preparation method, and belongs to the technical field of cell biology. Pancreas are rich in digestive enzymes, so that the treatment difficulty of in-vitro cells is high, and a high-quality single-cell suspension is difficult to prepare by a traditional method. The method comprises the steps of reagent preparation, tissue sampling, tissue dissociation, red cracking and cleaning, AOPI dyeing quality inspection and the like. Components and concentrations of a tissue enzymatic hydrolysate, a cell cleaning solution and a cell suspension are determined in reagent preparation; the acquisition method of the pancreatic tissue of the mouse is defined by tissue sampling; the tissue dissociation adopts a two-round digestion mode, and the cut tissue is subjected to enzymolysis by using a compound enzyme solution under specific conditions; the cell precipitates are treated through red cracking and cleaning; aOPI dyeing quality inspection is used for analyzing cell count and motility rate. According to the method, the culture medium containing specific components and the compound enzyme liquid are creatively used, the problem that pancreas single cells are difficult to dissociate can be effectively solved, the single-cell suspension with high motility rate, high purity and low adhesion is obtained, and the requirements of downstream single-cell experiments are met.

The application discloses a kind of high efficiency gentle mammal blastocyst single cell dissociation method.The method includes the following steps: (1) zona pellucida removal: the blastocyst is placed in calcium magnesium table type liquid, and zona pellucida is completely removed by dissolving and physical blowing operation;(2) enzyme digestion incubation: the blastocyst after step (1) is handled and is transferred to 35-37 ℃ pre-equilibrated complex dissociation solution and incubated for 15-30 minutes, during which intermittent blowing is carried out, until the blastocyst is completely dissociated into single cell;(3) washing: the single cell obtained by dissociation is washed using dPBS, and the residual dissociation solution is removed.The method of the application has high efficiency, high activity and simplified operation.The dissociation process is simplified from 3-4 steps to two steps (zona pellucida dissolution+enzyme digestion incubation), the total time is shortened from more than 90 minutes to less than 30 minutes, the dissociation efficiency is significantly improved;Cell viability is increased from about 70% to more than 90%, the cell junction is gently lysed, and cell damage is reduced.

The invention relates to the technical field of non-enzymatic cell dissociation, in particular to a non-enzymatic adipose tissue dissociation method and device based on hydrodynamic shearing, and the method comprises the following steps: step 1, obtaining dissociation indexes, and obtaining an optimal process parameter combination I meeting the dissociation indexes through a model based on the dissociation indexes; step 2, establishing a reference process based on the optimal process parameter combination I; step 3, judging whether a decision is input or not, if so, entering step 4, and otherwise, performing a dissociation process based on a reference process; and step 4, optimizing the reference process based on decision input to obtain an optimal process parameter combination 2, and performing a dissociation process based on the optimal process parameter combination 2.

The present application relates to the field of biotechnology, in particular to human bladder tissue digestion solution and application thereof.The digestion method of the present application can dissociate human bladder tissue cells while keeping the cell viability at more than 90%, and the amount of harvested cells is also obviously improved.Sufficient cell amount is the prerequisite for optimization treatment such as red blood cell lysis and dead cell removal.The single cell suspension obtained by the method has high viability, low clumping rate, high nucleated rate and less impurities, which can provide a basis for obtaining high-quality gene expression data in human bladder tissue single cell sequencing experiment.The present application can quickly obtain human bladder tissue primary cell suspension (sufficient cell amount, high viability, low clumping rate and less impurities) meeting the requirements of single cell experiment within 2-4 hours.

The present invention relates to new plasma or new platelet-rich plasma preparations, new cell dissociation methods, new cell associations or compositions, a method of preparation thereof, a use thereof, devices for the preparation thereof and preparations containing such a platelet-rich plasma preparation and cell associations or compositions. Specifically, the invention provides plasma or platelet-rich plasma alone or in cell composition preparations for use in tissue regeneration and bone regeneration and pain reduction.

This invention provides a kit, method, and single-cell library construction method for the dissociation of marine animal tissues. The dissociation method includes: S1) dissociating marine animal tissue samples at 8℃~12℃ using a cell dissociation buffer to obtain a single-cell suspension; S2) centrifuging the single-cell suspension and washing the single-cell precipitate obtained after centrifugation with a cell resuspension buffer to obtain single cells from the marine animal tissue sample; wherein, the cell dissociation buffer comprises a protease; the protease is Bacillus subtilis protease A; the concentration of sodium ions in the cell resuspension buffer is 300-800mM, and the concentration of potassium ions is 5-20mM.